Differentiation of Cirsium japonicum and C. setosum by TLC and HPLC-MS

The Chinese Pharmacopoeia indicates the use of field thistle (Cirsium setosum) and Japanese field thistle (C. japonicum) in the treatment of bleeding and inflammation. In the absence of an analytical method for the differentiation and analysis of these two species, TLC and HPLC-MS methods have been developed for this purpose. Both species could be readily distinguished by their flavonoid pattern as revealed by TLC on silica gel layers eluted with ethyl acetate:formic acid:acetic acid:water. The quantitative determination of four flavonoids, namely hispidulin-7-neohesperidoside, linarin, pectolinarin and luteolin, was possible using HPLC. Their optimum separation was achieved on a C12 column eluted with water and 0.025% trifluoroacetic acid in acetonitrile. HPLC-MS experiments were performed to confirm peak identity. In samples of C. japonicum, pectolinarin was the major flavonoid (0.32-2.00%), followed by linarin, hispidulin-7-neohesperidoside and luteolin; the total flavonoid content varied from 0.81 to 3.67%. In C. setosum only one flavonoid (linarin; 1.36-2.83%) was assignable. The HPLC method was validated for linearity, limit of detection (< or = 1.7 ng on-column), peak purity, repeatability (< or = 2.3%) and accuracy (recovery rates of spiked samples were between 99.2 and 101.6%).     

Cloning, expression and protective immunity evaluation of the full-length cDNA encoding succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum

1071-bp fragment was obtained from the Schistosoma japonicum (Chinese strain) adult cDNA library after the 3' and 5' ends of the incomplete expression sequence tag (EST) of succinate dehydrogenase iron-sulfur protein of Schistosoma japonicum (SjSDISP) were amplified by the anchored PCR with 2pairs of primers designed according to the EST of SjSDISP and the sequence of multiclone sites of the library vector. Sequence analysis indicated that the fragment was a full-length cDNA with a complete open reading frame (ORF), encoding 278 amino acid residues. The fragment was cloned into prokaryotic expression vector pQE30, and subsequently sequenced and expressed in Escherichia coll.SDS-PAGE and Western-blot analyses showed that the recombinant protein was about 32 kD and could be recognized by the polyclonal antisera from rabbits immunized with Schistosoma japonicum adult worm antigen. Compared with the FCA controls, mice vaccinated with rSjSDISP (test) or rSjGST (positive control) all revealed high levels of specific antibody and significant reduction in worm burden, liver eggs per gram (LEPG), fecal eggs per gram (FEPG) and intrauterine eggs. These results suggest that SjSDISP may be a novel and partially protective vaccine candidate against schistosomiasis. In contrast to the worm burden reduction rate, the higher degree of egg reduction rate in the test group also suggested that SjSDISP vaccine may primarily play a role in anti-embryonation or anti-fecundity immunity.     

Additional Notes on a Chrysosplenium L. (Saxifragaceae) from Anhui,China

A new variety Chrysosplenium japonicum var. cuneifolium from Anhui, China, is described, and the species Chrysosplenium delavayi is found new to Anhui. The genusChrysosplenium L. of Anhui is additionally recounted.     

海金沙草多糖的提取及抗氧化活性(英文)

用响应面优化技术研究了提取时间、固液比、提取温度等对海金沙草粗多糖提取的定量影响,获得了提取工艺的最优工艺参数:提取时间为123.3 min,固液比(s/w)为1∶20.9,提取温度为49.9℃,二阶多项式曲线回归模型预测多糖产量为12.466%,多糖提取验证试验结果(提取率)为12.85±0.18%(n=3),比模型预测稍高。多糖经纯化并进行体外抗氧化活性研究(以维生素C为对照品),结果发现,多糖产物对超氧阴离子(O2–.)和羟基氧自由基(.OH)具有较好的清除作用。     

Optimization of extracellular 5-aminolevulinic acid production from Escherichia coli transformed with ALA synthase gene of Bradyrhizobium japonicum

Extracellular accumulation of 5-aminolevulinic acid (ALA) by an E. coli overexpressing ALA synthase (ALAS) was achieved by inserting a hemA gene from Bradyrhizobium japonicum and expressed under the control of T7 promoter. At pH 7.0 extracellular ALA reached up to 15 mM in a jar fermenter with an addition of glycine (30 mM) and succinate (90 mM) in the medium. ALA accumulation was increased to 20 mM by adding levulinic acid (30 mM) to the cultures.     

安徽金腰属 (虎耳草科)植物新资料

楔叶金腰为金腰属一新变种,同时报道安徽新记录种肾萼金腰,并对安徽金腰属植物进行了补充记述。     

海金沙草总黄酮提取工艺的响应面优化

为了提高海金沙草总黄酮提取效率,运用了Plackett-Burman试验设计、爬陡坡试验和Box-Behnken设计对提取工艺进行了响应面优化试验。利用Plackett-Burman对影响总黄酮提取的诸多相关因素进行了评价并成功筛选出主效应因子,即提取时间、提取温度和乙醇浓度。在Plackett-Burman设计基础上,根据主效应因子作用大小与方向进行了爬陡坡试验。最后用Box-Behnken响应面技术优化了总黄酮提取工艺并建立了关键影响产量的二次多项式数学模型,解模型逆矩阵得最优解(优化方案),即提取温度X1=45.45℃,乙醇体积分数X2=47.1%、提取时间X3=84.8 min。模型预测结果为0.433 mg.L-1,验证试验结果为0.428±0.004 mg.L-1(n=3)。     

Photosynthetic rate and lectin activity of soybean leaves after inoculation with rhizobia together with homologous lectin

Nodule bacteria (Bradyrhizobium japonicum) of various activities were preincubated with homologous lectin and then used for inoculating soybean (Glycine max (L.) Merrill) seeds. The effect of this inoculation on the photosynthetic rate, lectin activity in leaves, and plant development at different supply of mineral nitrogen was investigated under the conditions of pot experiments. There was a positive relationship between the photosynthetic rate and the lectin activity of proteins isolated from soybean leaves. Under the conditions of effective symbiosis, activation of functioning of the symbiotic apparatus by preincubation of the rhizobia with lectin exerted an additional stimulating effect on the photosynthetic rate. It is suggested that a relationship between the effectiveness of legume-rhizobium symbiosis and the lectin activity in leaves is mediated by the regulation of photosynthesis through a demand for assimilates in the source-sink system of soybean plants.     

Fungi infecting cultivated moss can also cause diseases in crop plants

Bryophytes (mosses) are non‐vascular plants inhabited by a large number of fungal species, but whether mosses can act as reservoirs of fungal pathogens of crop plants has gained little attention. A few moss species including the Sunagoke moss (Racomitrium japonicum; family Grimmiaceae) are found to have modern economical applications in uses such as greening of urban environments. In a previous study, we identified fungi causing symptoms of varying severity in the commercially grown Sunagoke moss. The aim of this study was to test whether the same fungal isolates are pathogenic to vascular plants. An isolate of Fusarium avenaceum lethal to the Sunagoke moss caused root and crown rot in barley (Hordeum vulgare) and reduced germination of tomato (Solanum lycopersicum) and carrot (Daucus carota) grown in the infested soil. An isolate of Cladosporium oxysporum causing mild symptoms in moss reduced growth and caused reddening and premature death of carrot seedlings. On the other hand, isolates of Alternaria alternata and Fusarium oxysporum lethal to the Sunagoke moss caused no detectable symptoms in any tested vascular plant, suggesting specialisation of these isolates to moss. Chloroplast repositioning was observed in the neighbouring cells towards the initially infected cell following infection with F. avenaceum and A. alternata in Physcomitrella patens (family Funariaceae), a model moss used to study microscopic symptoms. Infection of P. patens with a non‐virulent Apiospora montagnei isolate induced formation of papillae in the moss cells, indicating activation of host defence as described in vascular plants. Results suggest that mosses and vascular plants may be linked by a common microbial interface constituted by pathogenic fungi. The findings have epidemiological implications that have gained little previous attention.     

肾上腺素受体和乙酰胆碱受体在血吸虫性肝纤维化肝脏组织中的表达研究

目的研究α1A-肾上腺素受体(α1A-AR)和M1型乙酰胆碱受体(mAChRM1)在日本血吸虫性肝纤维化小鼠肝脏组织中的表达情况,初步探讨神经递质受体在肝纤维化发生机制中的作用。方法清洁级昆明小鼠30只随机分为正常组(N组,10只)、模型组(M组,20只),模型组小鼠用腹部敷贴尾蚴法感染日本血吸虫制备肝纤维化模型。应用免疫组织化学、逆转录聚合酶链反应(RT-PCR)和免疫印迹(Western blot)等方法检测肝脏组织中α1A-AR和mAChRM1表达变化。结果模型组肝脏组织中α1A-AR和mAChRM1表达较正常组明显增加,其差异具有显著性统计学意义(P<0.05)。结论神经递质受体表达上调在血吸虫性肝纤维化发生发展中起一定作用。